Effect Of Sesamin On Expression Of JAK1 In Rabbit Aortic Endothelial Cells

Objective:This paper explores the role that sesamin plays in anti-atherosclerosis,on the basis of researches and comparisions on the effects of sesamin and AG490 on the expression of JAK1 in rabbit aortic endothelial cells during atherosclerosis.Methods:(1)Isolation,cell culture and passage of rabbit aortic endothelial cells.In the experiment,the logarithmic growth phase of aortic endothelial cells after successful passage is used to perfom(2)Cell grouping and treatment:①Normal control group:The experimental cells are cultured in serum-free DMEM medium for 24-48 hours under aseptic conditions;②ox-LDL group:The experimental cells are cultured in ox-LDL medium at a concentration of 100 mg/L for 24 hours and then in serum-free DMEM medium for 24 hours;③AG490(JAK1 inhibitor)group:The experimental cells are cultured in 50 μmol/L AG490 medium for 24 hours,then cultured in ox-LDL medium for 24 hours,and finally cultured in serum-free DMEM medium for 24 hours;④Sesamin group:The experimental cells are cultured in sesamin medium(60 μmol/L)for 24 hours,then cultured in 100 mg/L ox-LDL medium for 24 hours,and finally cultured in serum-free DEME medium 24 hour.(3)Real-time PCR is used to detect the effect of sesamin and AG490 on the expression of JAK1 in rabbit aortic endothelial cell proliferation.Results:In normal control group:the rabbit aortic endothelial cells whose shape is spindle or polygonal,the border is clear,rich in cytoplasm,and show a typical"paving stone" mosaic of cells,single distribution.In Ox-LDL group:the round rabbit aortic endothelial cells are soared,the border is unclear,the number is significantly reduced,apoptotic bodies appear,the cell debris increases,and the cell gap widens,occasionally falls off.In Schneinin group:the rabbit aortic endothelial cells whose shape is polygonal or spindle,the border was clear,the arrangement is close to the normal control group,but the number of cells is less than the normal control group and more than ox-LDL group.The PCR product is analyzed by 1%agarose gel electrophoresis and a specific band appeares at 250 bp.The JAK1 gene of rabbit aortic endothelial cells are successfully amplified.The recombinant plasmid is used as the template for the PCR reaction,and the upper and lower primers are used as primers.It shows that a specific band at 2945bp through the analysis by 1%agarose gel electrophoresis.The cloning plasmid of JAK1 gene of rabbit aortic endothelial cells are successfully amplified.The expression of JAKl-mRNA in ox-LDL group,sesamin group and AG490 group has been significantly higher than that in normal control group(P<0.05);The expression of JAK1-mRNA in sesamin group and AG490 group has been significantly lower than that in ox-LDL group(P<0.05);Sesamin and AG490 are significantly different(P<0.05).Conclusion:Both the saponin group and the AG490 group could inhibit the expression of JAK1-mRNA.The inhibitory effect of sesamin group is greater than that of JAK1 inhibitor AG490 group,can be used as a new target for prevention and treatment of cardiovascular and cerebrovascular diseases.